Evaluation of a <sub>D</sub>-Octaarginine-linked polymer as a transfection tool for transient and stable transgene expression in human and murine cell lines
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- SAKUMA Saki
- Laboratory of Immunology and Infection Control, Department of Veterinary Medicine, School of Veterinary Medicine, Azabu University, Kanagawa, Japan Present address: Division of Zoonosis Research, National Institute of Animal Health, National Agriculture and Food Research Organization, Ibaraki, Japan
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- OKAMOTO Mariko
- Laboratory of Immunology and Infection Control, Department of Veterinary Medicine, School of Veterinary Medicine, Azabu University, Kanagawa, Japan
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- MATSUSHITA Nao
- Faculty of Pharmaceutical Sciences, Setsunan University, Osaka, Japan
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- UKAWA Masami
- Faculty of Pharmaceutical Sciences, Setsunan University, Osaka, Japan
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- TOMONO Takumi
- Faculty of Pharmaceutical Sciences, Setsunan University, Osaka, Japan
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- KAWAMOTO Keiko
- Laboratory of Immunology and Infection Control, Department of Veterinary Medicine, School of Veterinary Medicine, Azabu University, Kanagawa, Japan
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- IKEDA Teruo
- Laboratory of Immunology and Infection Control, Department of Veterinary Medicine, School of Veterinary Medicine, Azabu University, Kanagawa, Japan
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- SAKUMA Shinji
- Faculty of Pharmaceutical Sciences, Setsunan University, Osaka, Japan
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Abstract
<p>Poly(N-vinylacetamide-co-acrylic acid) coupled with d-octaarginine (VP-R8) promotes the cellular uptake of peptides/proteins in vitro; however, details of the transfection efficacy of VP-R8, such as the cell types possessing high gene transfer, are not known. Herein, we compared the ability of VP-R8 to induce the cellular uptake of plasmid DNA in mouse and human cell lines from different tissues and organs. A green fluorescent protein (GFP)-expression plasmid was used as model genetic material, and fluorescence as an indicator of uptake and plasmid-derived protein expression. Three mouse and three human cell lines were incubated with a mixture of plasmid and VP-R8, and fluorescence analysis were performed two days after transfection. To confirm stable transgene expression, we performed drug selection three days after transfection. A commercially available polymer-based DNA transfection reagent (PTR) was used as the transfection control and standard for comparing transgene expression efficiency. In the case of transient transgene expression, slight-to-moderate GFP expression was observed in all cell lines transfected with plasmid via VP-R8; however, transfection efficiency was lower than using the PTR for gene delivery. In the case of stable transgene expression, VP-R8 promoted drug-resistance acquisition more efficiently than the PTR did. Cells that developed drug resistance after VP-R8-mediated gene transfection expressed GFP more efficiently than cells that developed drug resistance after transfection with the PTR. Thus, VP-R8 shows potential as an in vitro or ex vivo nonviral transfection tool for generating cell lines with stable transgene expression.</p>
Journal
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- Journal of Veterinary Medical Science
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Journal of Veterinary Medical Science 84 (4), 484-493, 2022
JAPANESE SOCIETY OF VETERINARY SCIENCE